The present invention relates to new substituted indolinones of general formula 
the isomers thereof, the salts thereof, particularly the physiologically acceptable salts thereof which have valuable properties.
The above compounds of general formula I wherein R4 denotes a hydrogen atom or a prodrug group have valuable pharmacological properties, in particular an inhibiting effect on various kinases, especially receptor tyrosine kinases such as VEGFR2, PDGFRxcex1, PDGFRxcex2, FGFR1, FGFR3, EGFR, HER2, IGF1R and HGFR, as well as complexes of CDK""s (Cycline Dependent Kinases) such as CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK8 and CDK9 with their specific cyclines (A, B1, B2, C, D1, D2, D3, E, F, G1, G2, H, I and K) and to viral cycline (cf. L. Mengtao in J. Virology 71 (3), 1984-1991 (1997)), to the proliferation of cultivated human cells, in particular endothelial cells, e.g. in angiogenesis, but also to the proliferation of other cells, in particular tumour cells.
The other compounds of the above general formula I wherein R4 does not denote a hydrogen atom or a prodrug group are valuable intermediate products for preparing the abovementioned compounds.
The present invention thus relates to the above compounds of general formula I, whilst those compounds wherein R4 denotes a hydrogen atom or a prodrug group have valuable pharmacological properties, pharmaceutical compositions containing the pharmacologically active compounds, the use thereof and processes for preparing them.
In the above general formula I
X denotes an oxygen or sulphur atom,
R1 denotes a C2-3-alkenyl, C2-3-alkynyl, aryl, aryl-C1-3-alkyl, heteroaryl, heteroaryl-C1-3-alkyl, trifluoromethyl or cyano group,
a hydroxy, C1-3-alkoxy, hydroxy-C1-3-alkyl, C1-3-alkoxy-C1-3-alkyl, aryloxy or heteroaryloxy group,
a mercapto, C1-3-alkylsulphenyl, phenylsulphenyl, benzylsulphenyl, C1-3-alkylsulphinyl, phenylsulphinyl, benzylsulphinyl, C1-3-alkylsulphonyl, phenylsulphonyl, benzylsulphonyl, sulpho, C1-3-alkoxysulphonyl, phenoxysulphonyl or benzyloxysulphonyl group,
an amino, C1-3-alkylamino, di-(C1-3-alkyl)-amino, hydroxycarbonyl-C1-3-alkylamino, Nxe2x80x94(C1-3-alkyl)-hydroxycarbonyl-C1-3-alkylamino, C1-3-alkoxycarbonyl-C1-3-alkylamino, Nxe2x80x94(C1-3-alkyl)-C1-3-alkoxycarbonyl-C1-3-alkylamino, phenylamino, N-phenyl-C1-3-alkylamino, N,N-diphenylamino, benzylamino, N-benzyl-C1-3-alkylamino, N,N-dibenzylamino, C1-3-alkylcarbonylamino, benzoylamino or benzylcarbonylamino group or an Nxe2x80x94(C1-3-alkyl)-C1-3-alkylcarbonylamino group wherein the two alkyl groups may be replaced by a C2-5-n-alkylene bridge or wherein one or both alkyl groups may be replaced by a phenyl or benzyl group,
a C1-3-alkylsulphonylamino, phenylsulphonylamino or benzylsulphonylamino group or an Nxe2x80x94(C1-3-alkyl)-C1-3-alkylsulphonylamino group wherein the two alkyl groups may be replaced by a C2-5-n-alkylene bridge or wherein one or both alkyl groups may be replaced by a phenyl or benzyl group,
an aminosulphonyl, C1-3-alkylaminosulphonyl, phenylaminosulphonyl, benzylaminosulphonyl, di-(C1-3-alkyl)-aminosulphonyl, oN,N-diphenyl-aminosulphonyl or N,N-dibenzyl-aminosulphonyl group,
a phosphono, (C1-3-alkoxy)PO(H), (C1-3-alkoxy)PO(C1-3-alkyl), (C1-3-alkoxy)PO(OH), di-(C1-3-alkoxy)-PO or (C2-4-n-alkylenedioxy)-PO group,
a ureido group optionally mono-, di- or trisubstituted by C1-3-alkyl groups,
a 4- to 7-membered cycloalkyleneimino or cycloalkyleneiminosulphonyl group, wherein in each case the methylene group in the 4 position of a 6- or 7-membered cycloalkyleneimino group may be replaced by an oxygen or sulphur atom, by a sulphinyl, sulphonyl, xe2x80x94NH or xe2x80x94N(C1-3-alkyl) group,
R2 denotes a hydrogen, fluorine, chlorine, bromine or iodine atom,
a C1-6-alkyl or trifluoromethyl group,
a hydroxy, C1-3-alkoxy, mercapto, C1-3-alkylsulphenyl, C1-3-alkylsulphinyl, C1-3-alkylsulphonyl, sulpho, C1-3-alkoxysulphonyl, aminosulphonyl, C1-3-alkylaminosulphonyl or di-(C1-3-alkyl)-aminosulphonyl group,
a nitro, amino, C1-3-alkylamino or di-(C1-3-alkyl)-amino group,
a C1-3-alkylcarbonyl, cyano, carboxy, C1-3-alkoxycarbonyl, aminocarbonyl, C1-3-alkylaminocarbonyl or di-(C1-3-alkyl)-aminocarbonyl group,
a phosphono, (C1-3-alkoxy)PO(H), (C1-3-alkoxy)PO(C1-3-alkyl), (C1-3-alkoxy)PO(OH) or di-(C1-3-alkoxy)-PO group,
a 4- to 7-membered cycloalkyleneimino, cycloalkyleneiminocarbonyl or cycloalkyleneiminosulphonyl group, wherein in each case the methylene group in the 4 position of a 6- or 7-membered cycloalkyleneimino group may be replaced by an oxygen or sulphur atom, by a sulphinyl, sulphonyl, xe2x80x94NH or xe2x80x94N(C1-3-alkyl) group, or
R1 and R2 together denote a methylenedioxy, ethylenedioxy, n-propylene, n-butylene or 1,4-butadienylene group,
R3 denotes a hydrogen atom, denotes a C1-6-alkyl, C3-7-cycloalkyl, trifluoromethyl or heteroaryl group,
a phenyl or naphthyl group optionally mono- or disubstituted by a fluorine, chlorine, bromine or iodine atom, by a trifluoromethyl, C1-3-alkyl or C1-3-alkoxy group, whilst in the case of disubstitution the substituents may be identical or different, which may additionally be substituted
by a hydroxy, hydroxy-C1-3-alkyl or C1-3-alkoxy-C1-3-alkyl group,
by a cyano, cyano-C1-3-alkyl, cyano-C2-3-alkenyl, cyano-C2-3-alkynyl, carboxy, carboxy-C1-3-alkyl, carboxy-C2-3-alkenyl, carboxy-C2-3-alkynyl, C1-3-alkoxycarbonyl, C1-3-alkoxycarbonyl-C1-3-alkyl, C1-3-alkoxycarbonyl-C2-3-alkenyl or C1-3-alkoxycarbonyl-C2-3-alkynyl group,
by a C1-3-alkylcarbonyl, C1-3-alkylcarbonyl-C1-3-alkyl, C1-3-alkylcarbonyl-C2-3-alkenyl or C1-3-alkylcarbonyl-C2-3-alkynyl group,
by an aminocarbonyl, aminocarbonyl-C1-3-alkyl, aminocarbonyl-C2-3-alkenyl, aminocarbonyl-C2-3-alkynyl, C1-3-alkylaminocarbonyl, C1-3-alkylaminocarbonyl-C1-3-alkyl, C1-3-alkylaminocarbonyl-C2-3-alkenyl, C1-3-alkylaminocarbonyl-C2-3-alkynyl, di-(C1-3-alkyl)-aminocarbonyl, di-(C1-3-alkyl)-aminocarbonyl-C1-3-alkyl, di-(C1-3-alkyl)-aminocarbonyl-C2-3-alkenyl or di-(C1-3-alkyl)-aminocarbonyl-C2-3-alkynyl group,
by a nitro, nitro-C1-3-alkyl, nitro-C2-3-alkenyl or nitro-C2-3-alkynyl group,
by an amino, C1-3-alkylamino, di-(C1-3-alkyl)-amino, amino-C1-3-alkyl, C1-3-alkylamino-C1-3-alkyl or di-(C1-3-alkyl)-amino-C1-3-alkyl group,
by a C1-3-alkylcarbonylamino, C1-3-alkylcarbonylamino-C1-3-alkyl, Nxe2x80x94(C1-3-alkyl)-C1-3-alkylcarbonylamino, Nxe2x80x94(C1-3-alkyl)-C1-3-alkylcarbonylamino-C1-3-alkyl, C1-3-alkylsulphonylamino, C1-3-alkylsulphonylamino-C1-3-alkyl, Nxe2x80x94(C1-3-alkyl)-C1-3-alkylsulphonylamino or Nxe2x80x94(C1-3-alkyl)-C1-3-alkylsulphonylamino-C1-3-alkyl group,
by a 4- to 7-membered cycloalkyleneimino, cycloalkyleneiminocarbonyl, cycloalkyleneiminosulphonyl, cycloalkyleneimino-C1-3-alkyl, cycloalkyleneiminocarbonyl-C1-3-alkyl or cycloalkyleneiminosulphonyl-C1-3-alkyl group, wherein in each case the methylene group in the 4 position of a 6- or 7-membered cycloalkyleneimino group may be replaced by an oxygen or sulphur atom, by a sulphinyl, sulphonyl, xe2x80x94NH or xe2x80x94N(C1-3-alkyl) group,
or by a heteroaryl or heteroaryl-C1-3-alkyl group,
R4 denotes a hydrogen atom, a C1-3-alkyl group or a prodrug group,
R5 denotes a hydrogen atom or a C1-3-alkyl group and
R6 denotes a hydrogen, fluorine, chlorine, bromine or iodine atom,
a trifluoromethyl or heteroaryl group, a C1-3-alkoxy group optionally substituted by 1 to 3 fluorine atoms, an amino-C1-3-alkoxy, C1-3-alkylamino-C2-3-alkoxy or benzylamino-C2-3-alkoxy group, a cycloalkyleneimino-C2-3-alkoxy group with 4 to 7 ring members, a di-(C1-3-alkyl)-amino-C2-3-alkoxy or C1-3-alkylmercapto group,
a nitro, cyano, carboxy, C1-3-alkoxycarbonyl, aminocarbonyl, C1-3-alkylaminocarbonyl, di-(C1-3-alkyl)-aminocarbonyl, piperidinocarbonyl or tetrazolyl group,
a C1-3-alkylcarbonylamino group optionally substituted at the nitrogen-atom by a C1-3-alkyl group,
an imidazolyl or piperazino group optionally substituted at the imino group by a C1-3-alkyl group,
a C1-4-alkyl group, which may be terminally substituted
by a hydroxy, C1-3-alkoxy, carboxy, C1-3-alkoxycarbonyl, amino, C1-4-alkylamino, di-(C1-4-alkyl)-amino, phenylamino, N-phenyl-C1-3-alkylamino, phenyl-n-C1-3-alkylamino, Nxe2x80x94(C1-3-alkyl)-phenyl-n-C1-3-alkylamino or di-(phenyl-n-C1-3-alkyl)-amino group,
by a 4- to 7-membered cycloalkyleneimino group wherein
a methylene group linked to the imino group may be replaced by a carbonyl or sulphonyl group or
one or two hydrogen atoms may each be replaced by a C1-3-alkyl group and/or
in each case the methylene group in the 4 position of a 6- or 7-membered cycloalkyleneimino group may be substituted by a carboxy, C1-3-alkoxycarbonyl, aminocarbonyl, C1-3-alkylaminocarbonyl, di-(C1-3-alkyl)-aminocarbonyl, phenyl-n-C1-3-alkylamino or Nxe2x80x94(C1-3-alkyl)-phenyl-n-C1-3-alkylamino group or may be replaced by an oxygen or sulphur atom, by a sulphinyl, sulphonyl, xe2x80x94NH or xe2x80x94N(C1-3-alkyl) group,
by a 5- to 7-membered cycloalkenyleneimino group wherein the double bond is isolated from the nitrogen atom,
by a C4-7-cycloalkylamino, Nxe2x80x94(C1-3-alkyl)-C4-7-cycloalkylamino or C5-7-cycloalkenylamino group wherein position 1 of the ring is not involved in the double bond and wherein the nitrogen atom may be substituted by a C1-3-alkyl group,
by a C1-3-alkylcarbonylamino, Nxe2x80x94(C1-3-alkyl)-C1-3-alkylcarbonylamino, aminocarbonyl, C1-3-alkylaminocarbonyl or di-(C1-3-alkyl)-aminocarbonyl group,
or R6 denotes a group of formula
xe2x80x94N(Ra)xe2x80x94COxe2x80x94(CH2)nxe2x80x94Rbxe2x80x83xe2x80x83(II),
xe2x80x83wherein
Ra denotes a C1-3-alkyl group,
n one of the numbers 0, 1 or 2 and
Rb denotes an amino, C1-4-alkylamino, phenylamino, Nxe2x80x94(C1-4-alkyl)-phenylamino, benzylamino, Nxe2x80x94(C1-4-alkyl)-benzylamino or di-(C1-4-alkyl)-amino group or a 4- to 7-membered cycloalkyleneimino group, wherein in each case the methylene group in the 4 position of a 6- or 7-membered cycloalkyleneimino group may be replaced by an oxygen or sulphur atom, by a sulphinyl, sulphonyl, xe2x80x94NH or xe2x80x94N(C1-3-alkyl) group,
a group of formula
xe2x80x94N(Rc)xe2x80x94(CH2)mxe2x80x94(CO)oxe2x80x94Rdxe2x80x83xe2x80x83(III),
xe2x80x83wherein
Rc denotes a C1-3-alkyl, C1-3-alkylcarbonyl, arylcarbonyl, benzylcarbonyl, C1-3-alkylsulphonyl, arylsulphonyl or benzylsulphonyl group,
m denotes one of the numbers 1, 2, 3 or 4,
o denotes one of the numbers 0 or 1 and
Rd has the meanings given for Rb hereinbefore or denotes a di-(C1-4-alkyl)-amino-C1-3-alkylamino group optionally substituted in the 1 position by a C1-3-alkyl group,
or an Nxe2x80x94(C1-3-alkyl)-C1-3-alkylsulphonylamino group,
whilst additionally any carboxy, amino or imino group present may be substituted by a group which can be cleaved in vivo (prodrug group),
and by a group which can be cleaved in vivo from an imino or amino group is meant, for example, a hydroxy group, an acyl group such as the benzoyl or pyridinoyl group or a C1-16-alkanoyl group such as the formyl, acetyl, propionyl, butanoyl, pentanoyl or hexanoyl group, an allyloxycarbonyl group, a C1-16-alkoxycarbonyl group such as the methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, tert.butoxycarbonyl, pentoxycarbonyl, hexyloxycarbonyl, octyloxycarbonyl, nonyloxycarbonyl, decyloxycarbonyl, undecyloxycarbonyl, dodecyloxycarbonyl or hexadecyloxycarbonyl group, a phenyl-C1-6-alkoxycarbonyl group such as the benzyloxycarbonyl, phenylethoxycarbonyl or phenylpropoxycarbonyl group, a C1-3-alkylsulphonyl-C2-4-alkoxycarbonyl, C1-3-alkoxy-C2-4-alkoxy-C2-4-alkoxycarbonyl or ReCOxe2x80x94Oxe2x80x94(RfCRg)xe2x80x94Oxe2x80x94CO group wherein
Re denotes a C1-8-alkyl, C5-7-cycloalkyl, phenyl or phenyl-C1-3-alkyl group,
Rf denotes a hydrogen atom, a C1-3-alkyl, C5-7-cycloalkyl or phenyl group and
Rg denotes a hydrogen atom, a C1-3-alkyl or ReCOxe2x80x94Oxe2x80x94(RfCRg)xe2x80x94O group wherein Re to Rg are as hereinbefore defined,
and additionally the phthalimido group may be used for an amino group, whilst the abovementioned ester groups may also be used as a group which can be converted in vivo into a carboxy group,
furthermore the term an aryl group denotes a phenyl or naphthyl group optionally mono- or disubstituted by a fluorine, chlorine, bromine or iodine atom or by a trifluoromethyl, C1-3-alkyl or C1-3-alkoxy group, whilst in the case of disubstitution the substituents may be identical or different, and
by a heteroaryl group is meant a monocyclic 5 or 6-membered heteroaryl group optionally substituted by one or two C1-3-alkyl groups, whilst the 6-membered heteroaryl group contains one, two or three nitrogen atoms and the 5-membered heteroaryl group contains an imino group optionally substituted by a C1-3-alkyl group, an oxygen or sulphur atom or an imino group optionally substituted by a C1-3-alkyl group and an oxygen or sulphur atom or one or two nitrogen atoms, and moreover a phenyl ring may be fused to the abovementioned monocyclic heterocyclic groups via two adjacent carbon atoms.
Particular mention should be made of the compounds of the abovementioned general formula I, wherein
X and R1 to R5 are as hereinbefore defined and
R6 is as hereinbefore defined, with the exception of an aminocarbonyl, C1-3-alkylaminocarbonyl, di-(C1-3-alkyl)-aminocarbonyl, piperidinocarbonyl or tetrazolyl group,
an imidazolyl or piperazino group optionally substituted by a C1-3-alkyl group at the imino group,
a C1-4-alkyl group, the terminal carboxy, C1-3-alkoxycarbonyl group,
an Nxe2x80x94(C1-3-alkyl)-C1-3-alkylsulphonylamino group
or a group of formula
xe2x80x94N(Rc)xe2x80x94(CH2)mxe2x80x94(CO)oxe2x80x94Rdxe2x80x83xe2x80x83(III),
xe2x80x83wherein
Rc denotes a C1-3-alkyl group.
Preferred compounds of the above general formula I are those wherein
X denotes an oxygen atom,
R1 denotes a C1-3-alkoxy, trifluoromethyl, di-(C1-3-alkyl)-amino, pyrrolidino or pyrrolo group,
an amino or C1-3-alkylamino group wherein an amino-hydrogen atom may be replaced by a C1-3-alkylcarbonyl, phenyl-C1-3-alkylcarbonyl, benzoyl, aminocarbonyl,
C1-3-alkylsulphonyl, phenylsulphonyl, carboxy-C1-3-alkyl or C1-3-alkyloxycarbonyl-C1-3-alkyl group, or
a phenyl group optionally substituted by a C1-3-alkyl group,
R2 denotes a hydrogen atom or a C1-3-alkoxy group or
R1 and R2 together denote a methylenedioxy group,
R3 denotes a C1-3-alkyl or phenyl group or a phenyl group substituted by a cyano, amino-C1-3-alkyl or Nxe2x80x94(C1-3-alkanoyl)-amino-C1-3-alkyl group,
R4 denotes a hydrogen atom,
R5 denotes a hydrogen atom and
R6 denotes a hydrogen, fluorine, chlorine, bromine or iodine atom,
a trifluoromethyl, 4-(C1-3-alkyl)-piperazino, pyridinyl, imidazolyl, tetrazolyl, C1-3-alkoxy or C1-3-alkylmercapto group,
a nitro, cyano, carboxy or C1-3-alkyloxycarbonyl group or a C1-3-alkylcarbonylamino group optionally substituted at the nitrogen atom by a C1-3-alkyl group,
a piperidinocarbonyl group or an aminocarbonyl group optionally substituted by one or two C1-3-alkyl groups,
a C1-3-alkyl group, which may be terminally substituted by an amino, C1-4-alkylamino, di-(C1-4-alkyl)-amino, phenylamino, N-phenyl-C1-3-alkylamino, phenyl-n-C1-3-alkylamino, Nxe2x80x94(C1-3-alkyl)-phenyl-n-C1-3-alkylamino or di-(phenyl-n-C1-3-alkyl)-amino group, by a pyrrolidino, piperidino, hexamethyleneimino, morpholino, thiomorpholino, 1-oxido-thiomorpholino or piperazino group,
whilst the piperidino group may additionally be substituted by one or two C1-3-alkyl groups or by a carboxy, C1-3-alkoxycarbonyl, aminocarbonyl, C1-3-alkylaminocarbonyl-di-(C1-3-alkyl)-aminocarbonyl or Nxe2x80x94(C1-3-alkyl)-phenyl-n-C1-3-alkylamino group,
by a C5-7-cycloalkylamino or C5-7-cycloalkenylamino group wherein position 1 of the ring is not involved in the double bond,
by a C1-3-alkylcarbonylamino, Nxe2x80x94(C1-3-alkyl)-C1-3-alkylcarbonylamino, carboxy, C1-3-alkoxycarbonyl, aminocarbonyl, C1-3-alkylaminocarbonyl or di-(C1-3-alkyl)-aminocarbonyl group,
a C1-3-alkoxy group, which is terminally substituted by an amino, C1-3-alkylamino or di-(C1-3-alkyl)-amino group,
a group of formula
xe2x80x94N(Ra)xe2x80x94COxe2x80x94(CH2)nxe2x80x94Rbxe2x80x83xe2x80x83(II),
xe2x80x83wherein
Ra denotes a C1-3-alkyl group,
n denotes one of the numbers 0, 1 or 2 and
Rb denotes an amino, C1-4-alkylamino or di-(C1-4-alkyl)-amino group or a pyrrolidino, piperidino, hexamethyleneimino, morpholino, thiomorpholino, 1-oxido-thiomorpholino or piperazino group,
a group of formula
xe2x80x94N(Rc)xe2x80x94(CH2)mxe2x80x94(CO)oxe2x80x94Rdxe2x80x83xe2x80x83(III),
xe2x80x83wherein
Rc denotes a C1-3-alkyl, C1-3-alkylcarbonyl or C1-3-alkylsulphonyl group,
m denotes one of the numbers 1, 2, 3 or 4,
o denotes one of the numbers 0 or 1 and
Rd has the meanings given for Rb hereinbefore or denotes a di-(C1-4-alkyl)-amino-C1-3-alkylamino group optionally substituted in the 1 position by a C1-3-alkyl group,
or an Nxe2x80x94(C1-3-alkyl)-C1-3-alkylsulphonylamino group,
the isomers and the salts thereof.
Particularly preferred compounds of the above general formula I are those wherein
X denotes an oxygen atom,
R1 denotes a methoxy, ethoxy, trifluoromethyl, phenyl, methylphenyl, dimethylamino, pyrrolidino or pyrrolo group, an amino group which may be substituted by a methyl, carboxymethyl, methoxycarbonylmethyl, acetyl, phenylacetyl, benzoyl, methanesulphonyl, benzolsulphonyl or aminocarbonyl group,
R2 denotes a hydrogen atom, a methoxy or ethoxy group or
R1 and R2 together denote a methylenedioxy group,
R3 denotes an ethyl group or a phenyl group optionally substituted by a cyano, aminomethyl or N-acetyl-aminomethyl group,
R4 denotes a hydrogen atom,
R5 denotes a hydrogen atom and
R6 denotes a hydrogen, fluorine, chlorine, bromine or iodine atom,
a methyl, trifluoromethyl, methoxy, ethoxy, methylmercapto, cyano, carboxy, methoxycarbonyl, ethoxycarbonyl, aminocarbonyl, dimethylaminocarbonyl, piperidinocarbonyl, nitro, 4-methyl-piperazino, imidazolyl, pyridinyl or tetrazolyl group,
an ethyloxy or n-propyloxy group terminally substituted by a dimethylamino group,
a methyl or ethyl group substituted by a carboxy, methoxycarbonyl, ethoxycarbonyl, aminocarbonyl or dimethylaminocarbonyl group,
a C1-3-alkyl group, which may be terminally substituted by an amino, C1-4-alkylamino, cyclohexylamino, benzylamino or phenylamino group wherein a hydrogen atom of the amino-nitrogen atom may be replaced in each case by a C1-3-alkyl, benzyl, acetyl or dimethylaminocarbonyl group,
by a piperidino group optionally substituted by one or two methyl groups,
by a piperidino group substituted by a carboxy, methoxycarbonyl, ethoxycarbonyl or dimethylaminocarbonyl group,
by a pyrrolidino, 3,4-dehydro-piperidino, hexamethyleneimino, morpholino, thiomorpholino, 1-oxo-thiomorpholino or piperazino group,
a C1-3-alkylamino group wherein the hydrogen atom of the amino-nitrogen atom is replaced
by an ethyl or n-propyl group, each of which is terminally substituted by a dimethylamino group,
by a C2-3-alkanoyl group which may be substituted in the 2 or 3 position by an amino, C1-3-alkylamino, di-(C1-3-alkyl)-amino, pyrrolidino, piperidino, morpholino or piperazino group,
by an aminocarbonyl, methylaminocarbonyl, dimethylaminocarbonyl, piperidinocarbonyl or methanesulphonyl group,
and additionally the C1-3-alkyl moiety of the C1-3-alkylamino group may be substituted
by an aminocarbonyl group,
by a C1-3-alkylaminocarbonyl or di-(C1-3-alkyl)-aminocarbonyl group wherein a C2-3-alkyl moiety may additionally be terminally substituted by a dimethylamino group, by a pyrrolidinocarbonyl, piperidinocarbonyl, morpholinocarbonyl or piperazinocarbonyl group,
whilst the C2-3-alkyl moiety of the abovementioned C1-3-alkylamino group may also be terminally substituted by an amino, C1-3-alkylamino, di-(C1-3-alkyl)-amino, pyrrolidino, piperidino, morpholino or piperazino group,
in particular those compounds of the above general formula I wherein
either X, R1 and R3 to R6 are as hereinbefore defined and R2 denotes a hydrogen atom,
or X and R3 to R6 are as hereinbefore defined, R1 and R2, which may be identical or different, each denotes a C1-3-alkoxy group,
the isomers and the salts thereof.
Quite specially preferred compounds of the above general formula I are those wherein
X denotes an oxygen atom,
R1 denotes an amino, methoxy or ethoxy group,
R2 denotes a hydrogen atom or in position 5 a methoxy or ethoxy group,
R3 denotes a methyl, ethyl or phenyl group,
R4 and R5 each denote a hydrogen atom and
R6 denotes a methyl or ethyl group substituted by a methylamino, ethylamino, piperidino or 4-(dimethylaminocarbonyl)-piperidino group, wherein the amino-hydrogen atom of the methylamino- and ethylamino group is replaced by a methyl or benzyl group, an N-dimethylaminomethylcarbonyl-N-methyl-amino group or an N-acetyl-Nxe2x80x94(C2-3-alkyl)-amino group wherein the C2-3-alkyl moiety in each case is terminally substituted by a dimethylamino group,
and the salts thereof.
The following particularly useful compounds of general formula I are mentioned by way of example;
(a) 3-(Z)-{1-[4-(piperidin-1-yl-methyl)-anilino]-1-phenyl-methylidene}-5,6-dimethoxy-2-indolinone,
(b) 3-(Z)-(1-{4-[(N-benzyl-N-methyl-amino)-methyl]-anilino}-1-phenyl-methylidene)-5,6-dimethoxy-2-indolinone,
(c) 3-(Z)-{1-(4-(dimethylamino-methyl)-anilino)-1-phenyl-methylidene}-5,6-dimethoxy-2-indolinone,
(d) 3-(Z)-{1-[4-(N-dimethylaminomethylcarbonyl-N-methyl-amino)-anilino]-1-phenyl-methylidene}-5,6-dimethoxy-2-indolinone,
(e) 3-(Z)-(1-{4-[2-(4-dimethylcarboxamide-piperidin-1-yl)-ethyl]-anilino}-1-phenyl-methylidene)-5,6-dimethoxy-2-indolinone,
(f) 3-(Z)-{1-[4-(N-dimethylaminomethylcarbonyl-N-methyl-amino)-anilino]-1-ethyl-methylidene}-5,6-dimethoxy-2-indolinone and
(g) 6-amino-3-(Z)-{1-[4-(piperidin-1-yl-methyl)-anilino]-1-phenyl-methylidene}-2-indolinone,
(h) 3-(Z)-(1-{4-[N-acetyl-N-(2-dimethylamino-ethyl)-amino]-anilino}-1-phenyl-methylidene)-5,6-dimethoxy-2-indolinone and
(i) 3-(Z)-(1-{4-[N-acetyl-N-(3-dimethylamino-propyl)-amino]-anilino}-1-phenyl-methylidene)-5,6-dimethoxy-2-indolinone
as well as the salts thereof.
According to the invention, the new compounds may be obtained, for example, by the following methods known in principle from the literature:
a. reaction of a compound of general formula 
wherein
X and R1 to R3 are defined as in claims 1 to 4,
R7 denotes a hydrogen atom, a protecting group for the nitrogen atom of the lactam group or a bond to a solid phase and
Z1 denotes a halogen atom, a hydroxy, alkoxy or arylalkoxy-group, e.g. a chlorine or bromine atom, a methoxy, ethoxy or benzyloxy group,
with an amine of general formula 
xe2x80x83wherein
R5 and R6 are defined as in claims 1 to 4, and if necessary subsequently cleaving any protecting group used for the nitrogen atom of the lactam group or cleaving from a solid phase.
A protecting group for the nitrogen atom of the lactam group might be for example an acetyl, benzoyl, ethoxycarbonyl, tert.butyloxycarbonyl or benzyloxycarbonyl group and
the solid phase might be a Rink resin such as a p-benzyloxybenzylalcohol resin, whilst the bond may conveniently be formed via an intermediate member such as a 2,5-dimethoxy-4-hydroxy-benzyl derivative.
The reaction is conveniently carried out in a solvent such as dimethylformamide, toluene, acetonitrile, tetrahydrofuran, dimethylsulphoxide, methylene chloride or mixtures thereof, optionally in the presence of an inert base such as triethylamine, N-ethyl-diisopropylamine or sodium hydrogen carbonate at temperatures between 20 and 175xc2x0 C., whilst any protecting group used can be cleaved simultaneously by transamidation.
If Z1 in a compound of general formula IV denotes a halogen atom, the reaction is preferably carried out in the presence of an inert base at temperatures between 20 and 120xc2x0 C.
If Z1 in a compound of general formula IV denotes a hydroxy, alkoxy or aralkoxy group, the reaction is preferably carried out at temperatures between 20 and 200xc2x0 C.
If any protecting group used subsequently has to be cleaved, this is conveniently carried out either hydrolytically in an aqueous or alcoholic solvent, e.g. in methanol/water, ethanol/water, isopropanol/water, tetrahydrofuran/water, dioxane/water, dimethylformamide/water, methanol or ethanol in the presence of an alkali metal base such as lithium hydroxide, sodium hydroxide or potassium hydroxide at temperatures between 0 and 100xc2x0 C., preferably at temperatures between 10 and 50xc2x0 C.,
or advantageously by transamidation with a primary or secondary organic base such as butylamine, dimethylamine or piperidine in a solvent such as methanol, ethanol, dimethylformamide and mixtures thereof or in an excess of the amine used at temperatures between 0 and 100xc2x0 C., preferably at temperatures between 10 and 50xc2x0 C.
Any solid phase used is preferably cleaved using trifluoroacetic acid and water in the presence of a dialkylsulphide such as dimethylsulphide at temperatures between 0 and 35xc2x0 C., preferably at ambient temperature.
b. In order to prepare a compound of general formula I wherein R1 denotes an amino group:
Reduction of a compound of general formula 
The reduction of a nitro group is preferably carried out by hydrogenolysis, e.g. with hydrogen in the presence of a catalyst such as palladium/charcoal or Raney nickel in a solvent such as methanol, ethanol, ethyl acetate, dimethylformamide, dimethylformamide/acetone or glacial acetic acid, optionally with the addition of an acid such as hydrochloric acid or glacial acetic acid at temperatures between 0 and 50xc2x0 C., but preferably at ambient temperature, and at a hydrogen pressure of 1 to 7 bar, but preferably 3 to 5 bar.
c. In order to prepare a compound of general formula I wherein R1 or/and R2 denotes one of the abovementioned substituted sulphinyl or sulphonyl groups:
oxidation of a compound of general formula 
xe2x80x83wherein
R3 to R6 are defined as in claims 1 to 4 and
one of the groups R1 and R2xe2x80x2 denotes one of the substituted mercapto or sulphinyl groups mentioned above for R1 and R2 and the other one assumes the meanings given above for R1 or R2 with the exception of the mercapto or sulphinyl groups or both groups R1xe2x80x2 and R2xe2x80x2, denote one of the substituted mercapto or sulphinyl groups mentioned above for R1 and R2.
The oxidation is preferably carried out in a solvent or mixture of solvents, e.g. in water, water/pyridine, acetone, methylene chloride, acetic acid, acetic acid/acetic anhydride, dilute sulphuric acid or trifluoroacetic acid, expediently at temperatures between xe2x88x9280 and 100xc2x0 C., depending on the oxidising agent used.
In order to prepare a corresponding sulphinyl compound of general formula I the oxidation is expediently carried out with one equivalent of the oxidising agent used, e.g. with hydrogen peroxide in glacial acetic acid, trifluoroacetic acid or formic acid at 0 to 20xc2x0 C. or in acetone at 0 to 60xc2x0 C., with a peracid such as performic acid in glacial acetic acid or trifluoroacetic acid at 0 to 50xc2x0 C. or with m-chloroperbenzoic acid in methylene chloride, chloroform or dioxane at xe2x88x9220 to 80xc2x0 C., with sodium metaperiodate in aqueous methanol or ethanol at xe2x88x9215 to 25xc2x0 C., with bromine in glacial acetic acid or aqueous acetic acid optionally in the presence of a weak base such as sodium acetate, with N-bromosuccinimide in ethanol, with tert.butylhypochlorite in methanol at xe2x88x9280 to xe2x88x9230xc2x0 C., with iodobenzodichloride in aqueous pyridine at 0 to 50xc2x0 C., with nitric acid in glacial acetic acid at 0 to 20xc2x0 C., with chromic acid in glacial acetic acid or in acetone at 0 to 20xc2x0 C. and with sulphurylchloride in methylene chloride at xe2x88x9270xc2x0 C., the resulting thioether-chlorine complex is expediently hydrolysed with aqueous ethanol.
In order to prepare a sulphonyl compound of general formula I the oxidation is expediently carried out starting from a corresponding sulphinyl compound with one or more equivalents of the oxidising agent used or starting from a corresponding mercapto compound, expediently with two or more equivalents of the oxidising agent used, e.g. with hydrogen peroxide in glacial acetic acid/acetic anhydride, trifluoroacetic acid or in formic acid at 20 to 100xc2x0 C. or in acetone at 0 to 60xc2x0 C., with a peracid such as performic acid or m-chloroperbenzoic acid in glacial acetic acid, trifluoroacetic acid, methylene chloride or chloroform at temperatures between 0 and 60xc2x0 C., with nitric acid in glacial acetic acid at 0 to 20xc2x0 C., with chromic acid, sodium periodate or potassium permanganate in acetic acid, water/sulphuric acid or in acetone at 0 to 20xc2x0 C.
If according to the invention a compound of general formula I is obtained which contains an alkoxycarbonyl group, this can be converted by hydrolysis into a corresponding carboxy compound, or
If a compound of general formula I is obtained which contains an amino or alkylamino group, this may be converted by alkylation or reductive alkylation into a corresponding alkylamino, dialkylamino or pyrrolidino compound, or
If a compound of general formula I is obtained which contains an amino or alkylamino group, this may be converted by acylation into a corresponding acyl compound, or
If a compound of general formula I is obtained which contains an amino or alkylamino group, this may be converted by sulphonation into a corresponding sulphonyl compound, for example into the corresponding alkylsulphonylamino, phenylsulphonylamino, benzylsulphonylamino or N-alkyl-alkylsulphonylamino group, or
If a compound of general formula I is obtained which contains an amino group, this can be converted by a reaction of condensation into a corresponding pyrrolo compound, or
If a compound of general formula I is obtained which contains a carboxy group, this may be converted by esterification or amidation into a corresponding ester or aminocarbonyl compound, or
If a compound of general formula I is obtained which contains a cyano group, this can be converted by reduction into a corresponding aminomethyl compound, or
If a compound of general formula I is obtained which contains an amino or alkylamino group, this may be converted by reaction with cyanic acid or a corresponding isocyanate into a corresponding ureido compound.
The subsequent hydrolysis is preferably carried out in an aqueous solvent, e.g. in water, methanol/water, ethanol/water, isopropanol/water, tetrahydrofuran/water or dioxane/water, in the presence of an acid such as trifluoroacetic acid, hydrochloric acid or sulphuric acid or in the presence of an alkali metal base such as lithium hydroxide, sodium hydroxide or potassium hydroxide at temperatures between 0 and 100xc2x0 C., preferably at temperatures between 10 and 50xc2x0 C.
The subsequent reductive alkylation is preferably carried out in a suitable solvent such as methanol, methanol/water, methanol/water/ammonia, ethanol, ether, tetrahydrofuran, dioxane, methylene chloride or dimethylformamide optionally with the addition of an acid such as hydrochloric acid in the presence of catalytically activated hydrogen, e.g. hydrogen in the presence of Raney nickel, platinum or palladium/charcoal, or in the presence of a metal hydride such as sodium borohydride, sodium cyanoborohydride, lithium borohydride or lithium aluminium hydride at temperatures between 0 and 100xc2x0 C., preferably at temperatures between 20 and 80xc2x0 C.
The subsequent alkylation is carried out with an alkylating agent such as an alkyl halide or dialkyl sulphate such as methyliodide, dimethylsulphate or propylbromide preferably in a solvent such as methanol, ethanol, methylene chloride, tetrahydrofuran, toluene, dioxane, dimethylsulphoxide or dimethylformamide optionally in the presence of an inorganic or a tertiary organic base such as potassium carbonate, triethylamine, N-ethyl-diisopropylamine, pyridine or dimethylaminopyridine, preferably at temperatures between 20xc2x0 C. and the boiling temperature of the solvent used.
The subsequent acylation is preferably carried out in a solvent such as methylene chloride, diethylether, tetrahydrofuran, toluene, dioxane, acetonitrile, dimethylsulphoxide or dimethylformamide, optionally in the presence of an inorganic or a tertiary organic base, preferably at temperatures between 20xc2x0 C. and the boiling temperature of the solvent used. The acylation with a corresponding acid is preferably carried out in the presence of a dehydrating agent, e.g. in the presence of isobutyl chloroformate, tetraethyl orthocarbonate, trimethyl orthoacetate, 2,2-dimethoxypropane, tetramethoxysilane, thionylchloride, trimethylchlorosilane, phosphorus trichloride, phosphorus pentoxide, N,Nxe2x80x2-dicyclohexylcarbodiimide, N,Nxe2x80x2-dicyclohexylcarbodiimide/N-hydroxysuccinimide, N,Nxe2x80x2-dicyclohexylcarbodiimide/1-hydroxy-benztriazole, 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium-tetrafluoroborate, 2-(1H-benzotriazol-1-yl)-1,1,3;3-tetramethyluronium-tetrafluoroborate/1-hydroxy-benzotriazole, N,Nxe2x80x2-carbonyldiimidazole or triphenylphosphine/carbon tetrachloride, and optionally with the addition of a base such as pyridine, 4-dimethylamino-pyridine, N-methyl-morpholine or triethylamine, conveniently at temperatures between 0 and 150xc2x0 C., preferably at temperatures between 0 and 100xc2x0 C., and the acylation with a corresponding reactive compound such as an anhydride, ester, imidazolide or halide thereof is optionally carried out in the presence of a tertiary organic base such as triethylamine, N-ethyl-diisopropylamine, N-methyl-morpholine or pyridine at temperatures between 0 and 150xc2x0 C., preferably at temperatures between 50 and 100xc2x0 C.
The subsequent sulphonation is preferably carried out in a solvent such as methylene chloride, diethylether, tetrahydrofuran, toluene, dioxane, acetonitrile, dimethylsulphoxide or dimethylformamide optionally in the presence of an inorganic or tertiary organic base, preferably at temperatures between 20xc2x0 C. and the boiling temperature of the solvent used. The sulphonation is carried out with a corresponding reactive compound, for example the sulphonylhalides, optionally in the presence of a tertiary organic base such as triethylamine, N-ethyl-diisopropylamine, N-methyl-morpholine, pyridine or dimethylaminopyridine at temperatures between 0 and 150xc2x0 C., preferably at temperatures between 50 and 100xc2x0 C.
The subsequent condensation for converting an amino group into a pyrrolo group is expediently carried out in a solvent such as dimethylformamide, toluene, acetonitrile, tetrahydrofuran, dimethylsulphoxide, methylene chloride or mixtures thereof, preferably in the presence of a acid such as formic acid, glacial acetic acid or trifluoroacetic acid or using the acid as the sole solvent with condesable 1,4-difunctional n-butylene or tetrahydrofuran derivatives, for example with 2,5-dimethoxytetrahydrofuran or 2,5-hexanedione, at temperatures between 0 and 120xc2x0 C. or the boiling temperature of the reaction mixture.
The subsequent esterification or amidation is expediently carried out by reacting a corresponding reactive carboxylic acid derivative with a corresponding alcohol or amine as described hereinbefore.
The subsequent reduction of a cyano group is preferably carried out in a suitable solvent such as methanol, methanol/water, methanol/water/ammonia, ethanol, ether, tetrahydrofuran, dioxane, methylene chloride or dimethylformamide optionally with the addition of methanolic ammonia with hydrogen in the presence of a hydrogenation catalyst, e.g. in the presence of Raney nickel or palladium/charcoal, at a hydrogen pressure of 1 to 5 bar, preferably at temperatures between 20xc2x0 C. and the boiling temperature of the solvent used.
The subsequent reaction with a cyanic acid or a corresponding isocyanate is preferably carried out in a suitable solvent such as ethanol, tetrahydrofuran or dioxane at temperatures between 0xc2x0 C. and the boiling temperature of the reaction mixture; the cyanic acid used is expediently prepared in the reaction mixture by reacting a salt of cyanic acid with an acid, for example by reacting potassium cyanate with glacial acetic acid.
In the reactions described hereinbefore, any reactive groups present such as carboxy, amino, alkylamino or imino groups may be protected during the reaction by conventional protecting groups which are cleaved again after the reaction.
For example, a protecting group for a carboxyl group may be a trimethylsilyl, methyl, ethyl, tert.butyl, benzyl or tetrahydropyranyl group and
protecting groups for an amino, alkylamino or imino group may be an acetyl, trifluoroacetyl, benzoyl, ethoxycarbonyl, tert.butoxycarbonyl, benzyloxycarbonyl, benzyl, methoxybenzyl or 2,4-dimethoxybenzyl group and additionally, for the amino group, a phthalyl group.
Any protecting group used is optionally subsequently cleaved for example by hydrolysis in an aqueous solvent, e.g. in water, isopropanol/water, tetrahydrofuran/water or dioxane/water, in the presence of a acid such as trifluoroacetic acid, hydrochloric acid or sulphuric acid or in the presence of an alkali metal base such as lithium hydroxide, sodium hydroxide or potassium hydroxide, at temperatures between 0 and 100xc2x0 C., preferably at temperatures between 10 and 50xc2x0 C.
However, a benzyl, methoxybenzyl or benzyloxycarbonyl group is cleaved, for example, hydrogenolytically, e.g. with hydrogen in the presence of a catalyst such as palladium/charcoal in a solvent such as methanol, ethanol, ethyl acetate, dimethylformamide, dimethylformamide/acetone or glacial acetic acid, optionally with the addition of an acid such as hydrochloric acid or glacial acetic acid at temperatures between 0 and 50xc2x0 C., but preferably at ambient temperature, and at a hydrogen pressure of 1 to 7 bar, but preferably 3 to 5 bar.
A methoxybenzyl group may also be cleaved in the presence of an oxidising agent such as cerium(IV)ammonium nitrate in a solvent such as methylene chloride, acetonitrile or acetonitrile/water at temperatures of between 0 and 50xc2x0 C., but preferably at ambient temperature.
A 2,4-dimethoxybenzyl group, however, is preferably cleaved in trifluoroacetic acid in the presence of anisol.
A tert.butyl or tert.butyloxycarbonyl group is preferably cleaved by treating with an acid such as trifluoroacetic acid or hydrochloric acid, optionally using a solvent such as methylene chloride, dioxane, ethyl acetate or ether.
A phthalyl group is preferably cleaved in the presence of hydrazine or a primary amine such as methylamine, ethylamine or n-butylamine in a solvent such as methanol, ethanol, isopropanol, toluene/water or dioxane at temperatures between 20 and 50xc2x0 C.
Moreover, chiral compounds of general formula I obtained may be resolved into their enantiomers and/or diastereomers.
Thus, for example, the compounds of general formula I obtained which occur as racemates may be separated by methods known per se (cf. Allinger N. L. and Eliel E. L. in xe2x80x9cTopics in Stereochemistryxe2x80x9d, Vol. 6, Wiley Interscience, 1971) into their optical antipodes and compounds of general formula I with at least 2 asymmetric carbon atoms may be resolved into their diastereomers on the basis of their physical-chemical differences using methods known per se, e.g. by chromatography and/or fractional crystallisation, and, if these compounds are obtained in racemic form, they may subsequently be resolved into the enantiomers as mentioned above.
The enantiomers are preferably separated by column separation on chiral phases or by recrystallisation from an optically active solvent or by reacting with an optically active substance which forms salts or derivatives such as e.g. esters or amides with the racemic compound, particularly acids and the activated derivatives or alcohols thereof, and separating the diastereomeric mixture of salts or derivatives thus obtained, e.g. on the basis of their differences in solubility, whilst the free antipodes may be released from the pure diastereomeric salts or derivatives by the action of suitable agents. Optically active acids in common use are e.g. the D- and L-forms of tartaric acid or dibenzoyltartaric acid, di-o-tolyltartaric acid, malic acid, mandelic acid, camphorsulphonic acid, glutamic acid, N-acetylglutamic acid, aspartic acid, N-acetylaspartic acid or quinic acid. An optically active alcohol may be for example (+)- or (xe2x88x92)-menthol and an optically active acyl group in amides, for example, may be a (+)- or (xe2x88x92)-menthyloxycarbonyl group.
Furthermore, the compounds of formula I obtained may be converted into the salts thereof, particularly for pharmaceutical use into the physiologically acceptable salts with inorganic or organic acids. Acids which may be used for this purpose include for example hydrochloric acid, hydrobromic acid, sulphuric acid, phosphoric acid, fumaric acid, succinic acid, lactic acid, citric acid, tartaric acid, maleic acid or methanesulphonic acid.
Moreover, if the new compounds of formula I thus obtained contain a carboxy group, they may subsequently, if desired, be converted into the salts thereof with inorganic or organic bases, particularly for pharmaceutical use into the physiologically acceptable salts thereof. Suitable bases for this purpose include for example sodium hydroxide, potassium hydroxide, cyclohexylamine, ethanolamine, diethanolamine and triethanolamine.
The compounds of general formulae IV to VII used as starting materials are known from the literature in some cases or may be obtained by methods known from the literature or are described in the Examples.
As already mentioned, the new compounds of general formula I wherein R4 denotes a hydrogen atom or a prodrug group have valuable pharmacological properties, particularly inhibitory effects on various kinases, especially on receptor-tyrosine kinases such as VEGFR2, PDGFRxcex1, PDGFRxcex2, FGFR1, FGFR3, EGFR, HER2, IGF1R and HGFR, as well as on complexes of CDK""s (Cycline Dependent Kinases) such as CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK8 and CDK9 with their specific cyclines (A, B1, B2, C, D1, D2, D3, E, F, G1, G2, H, I and K) and on viral cycline, on the proliferation of cultivated human cells, particularly endothelial cells, e.g. in angiogenesis, but also on the proliferation of other cells, particularly tumour cells.
The biological properties of the new compounds were tested by the following standard procedure, as follows:
Human umbilical endothelial cells (HUVEC) were cultivated in IMDM (Gibco BRL), supplemented with 10% foetal calf serum (FBS) (Sigma), 50 xcexcM of xcex2-mercaptoethanol (Fluka), standard antibiotics, 15 xcexcg/ml of endothelial cell growth factor (ECGS, Collaborative Biomedical Products) and 100 xcexcg/ml of heparin (Sigma) on gelatine-coated culture dishes (0.2% gelatine, Sigma) at 37xc2x0 C., under 5% CO2 in a water-saturated atmosphere.
In order to investigate the inhibitory activity of the compounds according to the invention the cells were xe2x80x9cstarvedxe2x80x9d for 16 hours, i.e. kept in culture medium without growth factors (ECGS+heparin). The cells were detached from the culture dishes using trypsin/EDTA and washed once in serum-containing medium. Then they were seeded out in amounts of 2.5xc3x97103 cells per well.
The proliferation of the cells was stimulated with 5 ng/ml of VEGF165 (vascular endothelial growth factor; H. Weich, GBF Braunschweig) and 10 xcexcg/ml of heparin. As a control, 6 wells in each dish were not stimulated.
The compounds according to the invention were dissolved in 100% dimethylsulphoxide and added to the cultures in various dilutions as triple measurements, the maximum dimethyl sulphoxide concentration being 0.3%.
The cells were incubated for 76 hours at 37xc2x0 C., then for a further 16 hours 3H-thymidine (0.1 xcexcCi/well, Amersham) was added in order to determine the DNA synthesis. Then the radioactively labelled cells were immobilised on filter mats and the radioactivity incorporated was measured in a xcex2-counter. In order to determine the inhibitory activity of the compounds according to the invention the mean value of the non-stimulated cells was subtracted from the mean value of the factor-stimulated cells (in the presence or absence of the compounds according to the invention).
The relative cell proliferation was calculated as a percentage of the control (HUVEC without inhibitor) and the concentration of active substance which inhibits the proliferation of the cells by 50% (IC50) was determined.
The following Table contains the results found:
In addition the compound of Example 1 (6) was tested on mice for its tolerance. The approximate oral LD50 for this compound is over 1,000 mg/kg. The compounds of general formula I are therefore well tolerated.
In view of their inhibitory effect on the proliferation of cells, particularly endothelial cells and tumour cells, the compounds of general formula I are suitable for treating diseases in which the proliferation of cells, particularly endothelial cells, plays a part.
Thus, for example, the proliferation of endothelial cells and the concomitant neovascularisation constitute a crucial stage in tumour progression (Folkman J. et al., Nature 339, 58-61, (1989); Hanahan D. and Folkman J., Cell 86, 353-365, (1996)). Furthermore, the proliferation of endothelial cells is also important in haemangiomas, in metastasisation, rheumatoid arthritis, psoriasis and ocular neovascularisation (Folkman J., Nature Med. 1, 27-31, (1995)). The therapeutic usefulness of inhibitors of endothelial cell proliferation was demonstrated in the animal model for example by O""Reilly et al. and Parangi et al. (O""Reilly M. S. et al., Cell 88, 277-285, (1997); Parangi S. et al., Proc Natl Acad Sci USA 93, 2002-2007, (1996)).
The compounds of general formula I, their tautomers, their stereoisomers or the physiologically acceptable salts thereof are thus suitable, for example, for treating solid tumours, diabetic retinopathy, rheumatoid arthritis and psoriasis, or other diseases in which cell proliferation or angiogenesis play a part.
By reason of their biological properties the compounds according to the invention may be used on their own or in conjunction with other pharmacologically active compounds, for example in tumour therapy, in monotherapy or in conjunction with other anti-tumour therapeutic agents, for example in combination with Topoisomerase inhibitors (e.g. Etoposide), mitosis inhibitors (e.g. Vinblastin, Taxol), compounds which interact with nucleic acids (e.g. cisplatin, Cyclophosphamide, Adriamycin), hormone antagonists (e.g. tamoxifen), inhibitors of metabolic processes (e.g. 5-FU etc.), cytokines (e.g. interferons), kinase inhibitos, antibodies, or in conjunction with radiotherapy, etc. These combinations may be administered either simultaneously or sequentially.
For pharmaceutical use the compounds according to the invention are generally used for warm-blooded vertebrates, particularly humans, in doses of 0.01-100 mg/kg of body weight, preferably 0.1-15 mg/kg. For administration they are formulated with one or more conventional inert carriers and/or diluents, e.g. with corn starch, lactose, glucose, microcrystalline cellulose, magnesium stearate, polyvinylpyrrolidone, citric acid, tartaric acid, water, water/ethanol, water/glycerol, water/sorbitol, water/polyethyleneglycol, propyleneglycol, stearylalcohol, carboxymethylcellulose or fatty substances such as hard fat or suitable mixtures thereof in conventional galenic preparations such as plain or coated tablets, capsules, powders, suspensions, solutions, sprays or suppositories.
The following Examples are intended to illustrate the present invention without restricting it:
Abbreviations:
TBTU=O-(benzotriazol-1-yl)-N,N,Nxe2x80x2,Nxe2x80x2-bis(tetramethylene)-uronium hexafluorophosphate
HOBt=1-hydroxy-1H-benzotriazole
Preparation of the starting compounds: